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2.79J/3.96J/20.441J/HST522JBiomaterials-Tissue Interactions
Structure and function of naturally occurring extracellular
matrices (ECMs).
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The insoluble regulator in a unit cell process is part of an ECM macromolecule
Cell + InsolubleRegulator Product
SolubleRegulator A
SolubleRegulator B
Control volume dV
Unit cell process confined conceptually in a control volume dV
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Integrin adhesion to collagen at the GFOGER ligand
Emsley et al., 2000Knight et al., 2000
Phenotype change: fibroblast α2β1 integrin binds to collagen ligand GFOGER (hexapeptide)
Courtesy of Elsevier, Inc., http://www.sciencedirect.com.Used with permission.
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The Extracellular Matrices (ECMs) (summary of structure and function)
Insoluble macromolecular networks. Structure varies with organ; but different ECMs comprise few types of macromolecules (mostly collagen, elastin, proteoglycans) plus water (65%).ECM does not migrate, proliferate, synthesize proteins or contain DNA!Give and take of signals with cells. Ligands on ECM surface interact specifically with cell receptors (integrins). Partly determine the state of differentiation of cells.
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The Extracellular Matrices…(summary of structure and function)
Possibly play role of memory storage device which is used to record events (e.g., a recent cell migration), thereby informing cells of what has already been done and acting as “arrow” in a kinetic process. Often bind cytokines and growth factors and act as reservoirs of such molecules.Loss of cell-matrix contact characterizes tumor cells just prior to spreading of cancer from one organ to another (metastasis).Determines the shape of animals and maintains positional homeostasis of organs.Recently, certain synthetic ECM models have induced organ regeneration in adults.
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The major ECM molecules in tissues
1. Collagens.2. Elastin.3. Proteoglycans and glycosaminoglycans (GAGs).4. Cell-adhesion molecules (fibronectin, laminin,
others).[Water (about 65% of tissue weight).]
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Hierarchy of structural order in proteins
Primary structure: the complete sequence of amino acids (AA) in the polypeptide chain. Scale: 1 nm. Secondary structure: the local chain configuration (sequence of 3 - 5 AA). Scale: 10 nm. Tertiary structure: the configuration of the entire macromolecule. Scale: 100 nm.
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Hierarchy of structural order in proteins (cont.)
Quaternary structure: The packing pattern of several identical molecules that characterizes a crystalline fiber. Scale: 1000 nm = 1 μm.
Architecture: Pattern comprising several fibers of a protein that constitute a macroscopic tissue. Often contains fibers of two different proteins (collagen and elastin) and one or more proteoglycan molecules. Scale: 1-10 mm.
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Collagens(most fibrous collagens)
Primary structure: Glycine “hinge” every third AA makes polypeptide chains capable of rotation. Hydroxyproline (25% of total AA content) stiffens polypeptide chains. Varies with organ; several such “collagens” have been identified. Fibrous collagens to be discussed here only.
Secondary structure: Combination of hinge-like glycine and stiff hydroxyproline units, leads to helical macromolecule with sharp pitch.
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Tertiary structure
Collagens (cont.)(most fibrous collagens)
: Three helical polypeptide units twist to form a triple-helical collagen molecule: a molecular “rope” which has some bending stiffness and does not undergo rotation.
Quaternary structure: Several collagen molecules pack side-by-side in a highly specific register to give a crystalline fiber with a 64-nm periodicity (collagen banding pattern).
The architectural structure of collagen is uniaxial orientation in tendon, biaxial orientation in the dermis, etc. It determines the mechanical behavior of the tissue.
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COLLAGEN STRUCTURE
Primary
Secondary
Tertiary
Quaternary banding
Diagram removed due to copyright restrictions.
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Cross-linking of collagen molecules
Diagram removed due to copyright restrictions.Formation of intramolecular and intermolecular cross-links in type I collagen.
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Collagen structure-function relations
• The primary structure of collagen is tissue-specific. Type I in tendon, type II in cartilage, etc.
• The secondary and the tertiary structures are specific substrates for the metalloprotein enzyme collagenase that degrades collagen fibers. Remodeling of tissues during wound healing by collagenase. Melting of collagen to gelatin (loss of tertiary structure) spontaneously follows such degradation.
• The banding (quaternary structure) of collagen fibers determines the blood clot-forming properties of collagen (primarily through induction of platelet aggregation).
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Collagen structure-function relations (cont.)
• The architectural structure of collagen determines the function of collagen fibers as mechanical reinforcements of connective tissues (tendon, skin, bone, arteries etc.). Tendon fibers are bundles of uniaxially aligned fibers that are crimped. Skin (dermis) is a random planar array of crimped collagen fibers. Bone is a ductile ceramic (hydroxyapatite) which is reinforced by collagen fibers. Large blood vessels (aorta, large arteries) are interpenetrating networks of elastin fibers and collagen fibers.
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Exposed to collagenase
No enzyme
Exposed longer
Effect of exposure of collagen fibers to collagenase
Three photos removed due to copyright restrictions.
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Degradation of collagen molecule by collagenase(Gross)
spontaneous melting to gelatin following degradation
collagen
Diagram removed due to copyright restrictions.
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Cleavage site of collagen molecule by collagenase is within the fibronectin binding region (766-786)
Diagram removed due to copyright restrictions.
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The Extracellular Matrices Part II.
2. Elastin fibers.3. Proteoglycans (PG) and glycosaminoglycans
(GAG).4. Cell-adhesion molecules (CAM).
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Elastin fibers• A network of randomly coiled macromolecules.
No periodicity. Highly extensible chains. • Rubber-like elasticity is complicated by
hydrophobic bonding effects.• Interaction of hydrophobic (nonpolar) AA with
water leads to hydrophobic bonding. Primarily entropic, not energetic, bonding between molecules. It forces nonpolar macromolecules, such as elastin, to adopt a compact, rather then extended, shape in hydrated tissue.
• Stretching of elastin fibers leads to large entropy loss due to reduction in chain configurations and increased “ordering” of water molecules against nonpolar AA. Spontaneous retraction.
• Elastic ligament of neck. Blood vessel wall.
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The Hydrophobic bondΔG = ΔH − TΔS
Equilibrium when ΔG = 0. G is Gibbs’ free energy, the enthalpy is H = E + PV, T is absolute temperature andS is the entropy. The process goes spontaneously from left to right when ΔG < 0. Find the position of thermodynamic equilibrium for a well-known exampleof insolubility:
CH4 in benzene → CH4 in H2OThe experimental data show (all units in calories per mol): ΔG = Δ H − T Δ S
+2600 = −2800 − 298(−18)+2600 = −2800 + 5400
Conclusion: Insolubility of paraffin in water due to entropy loss, not to enthalpy change! (Kauzmann)
26Historical models of cell membranestructure
Diagram removed due to copyright restrictions.Models of (a) Goldup, Ohki and Danielli (b) Singer and Nicolson (c) Lenard and Singer (d) Lucy (e) Kreutz (f) Vanderkooi and Green.
Cell membraneshowing bilayer structure
Figure by MIT OpenCourseWare.
Figure by MIT OpenCourseWare.Figure by MIT OpenCourseWare.
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Macromolecules coil upon themselves due tohigh content of nonpolar (hydrophobic) amino acids that mediate withdrawal from polar medium (aqueous buffer) and promote bonding within chains.These networksstretch extensively like all rubbers.
Elastin fibers in the relaxed aorta. Elastin macromolecules are random coils tied together to forma 3-dimensional (insoluble) network.
Two histology photos removed due to copyright restrictions.
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Proteoglycans (PGs) and glycosaminoglycans (GAGs)
• A proteoglycan is a polypeptide chain (proteo) with polysaccharide (glycan or GAG) side chains.
• Primary structure modeled as an alternating copolymer of two different glucose-like units, one of them an acidic sugar-like molecule, the other an amino sugar with a negatively charged sulfate group (except hyaluronic acid that is not sulfated).
• Electrostatic interactions between charged groups in GAG side chains of PG responsible for about 50% of stiffness of articular cartilage (Grodzinsky et al.).
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Proteoglycans (PGs) and glycosaminoglycans (GAGs)
GAG side chains of PG
PG
PG
Diagram removed due to copyright restrictions.Structures of four subpopulations of cartilage proteoglycans.
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Proteoglycans and glycosaminoglycans
repeat unit of chondroitin 6-sulfate
a proteoglycan
Diagram removed due to copyright restrictions.
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Electron microscopic view of a proteoglycanfrom bovine nasal cartilage
Image removed due to copyright restrictions.
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Cell-adhesion molecules
• Cell-matrix interactions involve binding of transmembrane proteins (integrins) on specific sites (ligands) in ECM molecules such as fibronectin, laminin and collagen. Example: integrins of contractile fibroblasts bind to fibronectin molecules that are attached to collagen fibers (fibronexus).
• Integrins connect with proteins in the cell cytoplasm and a signal is transmitted to or from the nucleus.
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Μια ιντεγκρινη συνδεει το εσωτερικο του κυτταρου (κατω) με ΕΚΜ (πανω)
An integrin “connects” the interior of the cell (cytoplasm) with the ECM outside it
Figure from (Hynes, 1990)
cell membrane
Figure by MIT OpenCourseWare.
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Cell membraneshowing cell surface receptors (integrins)
Phospholipidmolecule
Bilayer structure of cell membraneviewed by electron microscopy
Figure by MIT OpenCourseWare.
Figure by MIT OpenCourseWare. Figure by MIT OpenCourseWare.
MIT OpenCourseWarehttp://ocw.mit.edu
20.441J / 2.79J / 3.96J / HST.522J Biomaterials-Tissue InteractionsFall 2009
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